Reverse Transcriptase PCR (RT PCR)

One of the best features of a particular cellular function state is its gene expression pattern. Cells belonging to different tissues, cells at different stages of development or metabolism, cells affected by a particular compound or cells in an oncogenic process differ in their mRNA libraries due to their gene expression pattern. Presently, the most important technique for accurate quantification of gene expression is fluorometric real-time RT-PCR (Muller et al., 2002a). Reverse transcription (RT) followed by the polymerase chain reaction (PCR) is a selection technique for analyzing mRNA expression from various sources.

In recent years reverse transcriptase PCR (RT-PCR) has been established for digital RNA viruses, which can modernize laboratory diagnosis of dengue fever (28). RT-PCR provides rapid diagnosis of specific sera. If properly handled, this method is rapid, simple, sensitive and reproducible and can be used to detect viral RNA in human clinical specimens and autopsy tissues or mosquitoes. In recent years, there are many ways to use primer genomes at different locations and different methods to detect RT-PCR products (29).

When transcribing the original sample of RNA reverse transcriptase PCR (RT - PCR) is used so that the DNA is the product of amplification. The detection comes from a single nucleotide base and the sensitivity of PCR is high because its quantitative capability comes from amplification of the amplified DNA to its original size. Southern blots begin with DNA samples that are initially degraded into smaller, altered fragments by restriction endonucleases. The DNA is then placed in wells for agarose gel electrophoresis, where the fragments diffuse through the polarization field according to their size. The DNA was denatured with sodium hydroxide, transferred to nitrocellulose or nylon sheets and incubated with single stranded DNA hybridization probe. The radiolabelled probe binds to the exposed complementary base pair and can be detected by autoradiography

Inspection of mosquito samples requires direct amplification using reverse transcriptase PCR (RT-PCR) and indicates the presence of virus in the submitted samples. When using serum of wild birds or sentinel chickens, it is necessary to test for the presence of WNV antibodies in the sample using immunohistochemistry (IHC) or enzyme-linked immunosorbent assay (ELISA). Environmentalists condemn the attempt to prevent the spread of mosquitoes by spraying insecticides and the harmful effects of spraying on health is greater than the relatively small life that can be saved and mosquitoes Said it could be controlled in a more environmentally friendly way. They also question the effectiveness of pesticide spray as they believe that resting beyond spray level or flying mosquitoes will not be killed; the most common carrier in the northeastern United States, Culex pipiens Is a canopy feeder.